Production of L-isoleucine by means of recombinant microorganisms with deregulated threonine dehydratase

ABSTRACT

The invention relates to processes for the microbial production of L-isoleucine. To this end, in a gene in vitro of a threonine dehydratse, one or more bases in the gene region coding the enzyme&#39;s allosteric domains are exchanged in such a way that at least one amino acid in the amino acid sequence of the allosteric domains of the enzyme is replaced by another so that the enzyme is no longer inhibited by L-isoleucine feedback. Furthermore, concrete amino acid exchanges in the amino acid sequence of the enzyme are effected in a gene in vitro of a threonin dehydratase of Corynebacterium glutamicum by base exchange both outside and inside and outside the gene region coding the allosteric domains of the enzyme si that, after the transformation of such mutated threonine dehydratase genes into a threonine or L-isoleucine-producing host cell, the latter repeatedly forms L-isoleucine.

This application is a National Phase filing of PCT/DE95/00017, filedJan. 9, 1995.

The invention relates to a method for the microbial production ofL-isoleucine wherein a threonine dehydratase gene is altered so as toderegulate the feed back inhibition of L-isoleucine by threoninedehydratase. The invention also relates to a gene structure and vectorcorresponding to the altered threonine dehydratase gene. The inventionalso relates to a transformed cell which contains the aforementionedgene structure.

The amino acid L-isoleucine is essential for man and animals. It iswidely used in dietetic foods and as a component of various nutrientmixtures for medicinal purposes. L-Isoleucine is also used as anadditive or as a reagent in the pharmaceutical and chemical industries.

Microorganisms, which secrete L-isoleucine into a fermentation medium,are used to obtain this amino acid, which is formed by the biosyntheticpath shown in FIG. 1. As is also demonstrated in FIG. 1, there are keyenzymes in the biosynthesis of the L-isoleucine, namely aspartic acidkinase, homoserine dehydrogenase and threonine dehydratase. The activityof the aspartic acid kinase and of the homoserine dehydrogenase isinhibited by the feedback of the amino acid, L-threonine, which is alsoformed within the scope of the biosynthesis of the L-isoleucine, whilethe key enzyme, threonine dehydratase, which is specific for thesynthesis of L-isoleucine, is inhibited by the feedback of the endproduct of the biosynthetic chain, L-isoleucine.

In the past, attempts have been made time and again to increase theformation of L-isoleucine by obtaining mutants of L-isoleucine producerswhich, relative to the wild types, form more L-isoleucine.

To obtain such mutants, mutagenesis was carried out exclusively in vivo,that is, a mutagen was allowed to act on the whole of the genome.Mutated microorganisms, the above-mentioned key enzymes of which nolonger are subject to a feed-back inhibition, were selected on the basisof the resistance to amino acid analogs.

For example, a mutation, which leads to a resistance to α-aminobutyrateor isoleucine hydroxamate, is described in U.S. Pat. No. 4,329,427.According to this example, microorganisms admittedly produce increasedamounts of isoleucine; however, the question as to which enzymes are nolonger inhibited by feedback, is not clarified.

The U.S. Pat. No. 4,442,208 and 4,601,983 disclose that, after an invivo mutagenesis, it was possible to isolate a DNA fragment, which isnot defined more precisely and which brings about resistance toα-aminohydroxyvaleric acid. After this fragment is transferred to aCorynebacterium or a Brevibacterium strain, the latter produceincreasing amounts of L-isoleucine.

Methods have also been described, for which, by means of oversynthesisof feedback-regulated key enzymes, L-isoleucine can be formedincreasingly (see, for example, the German Offenlegungsschrift 3,942,947and the European publication 0 137 348).

Overall, all previously described methods for increasing theL-isoleucine production have the common feature that mutations were morelikely to lead randomly to deregulated key enzymes, which are no longersubject to feedback inhibition and that the amino acid synthesis wasincreased by these means.

It is an object of the invention to provide a method for the microbialproduction of L-isoleucine, by means of which defined mutations of thekey enzyme of the L-isoleucine biosynthesis, the threonine dehydratase,is selectively changed so that feedback inhibition by L-isoleucine nolonger takes place.

The objective, on which the invention is based, is accomplished owing tothe fact that, in a gene of a threonine dehydratase, present in vitro,one or several bases are exchanged in such a manner by mutation in theregion of the gene coding for the allosteric domains of the enzyme, thatat least one amino acid in the amino acid sequence of the allostericdomains of the enzyme is replaced by a different one in such a manner,that the enzyme is no longer inhibited by L-isoleucine feedback. "Agene, present in vitro" in this case means that the threoninedehydratase gene is cloned, that is, isolated and incorporated in avector, before mutagenesis. The mutagenesis by exchange of base(s) isaccomplished by known methods, for example, by the method of Ito et al.(Gene 102 (1991) 67-70). After transformation of threonine dehydratasegenes, so mutated, in a host cell producing threonine or L-isoleucine,the host cell increasingly produces L-isoleucine.

In principle, a cloned threonine dehydratase gene or one present invitro can originate from any bacterium. Since Corynebacterium glutamicumis one of the "classical" amino acid producers, the gene preferablyoriginates from this microorganism, particularly from the ATCC 13032strain. The DNA sequence of the threonine dehydratase gene from thisCorynebacterium glutamicum strain is already known (see Mockel et al.,J. Bacteriol. 174 (1992) 8065-8072). After a base exchange in the generegion coding for the allosteric domains of the enzyme, the DNAsequences, listed in Tables 1 to 3, (SEQ. ID. Nos. 1, 3 & 5,respectively) are obtained for example. In the Tables, the gene region,coding for the allosteric domains of the enzymes, is underlined and thebase exchange sites are indicated by additional underlining.

After the mutagenesis is carried out, the in vitro mutated genes areincorporated in a plasmid, in which the expression of the threoninedehydratase gene can be induced. As plasmids, pVC 19 or pKK 223-3, forexample, are suitable (Amman et al., Gene 25, 167). After incorporationof the mutated genes in a suitable plasmid, they are transformed in asuitable bacterial strain.

In order to obtain those clones, which also actually contain theplasmids with the mutated threonine dehydratase genes, the transformantsdesired can be isolated by the usual method by way of the resistance ofamino acid analogs, β-methylnorleucine, isoleucine hydroxamate orhydroxyisoleucine coming into consideration as analogs.

However, a simpler and more concerted isolation is attained owing to thefact that the changed threonine dehydratase genes are transformed in amicroorganism, the acetohydroxy acid synthase activity of which can beinhibited by L-valine. Escherichia coli K 12 strains, preferably thestrains JM109 of DH 5, are suitable as transformants. The transformantsare subsequently brought onto a solid medium, which contains theL-isoleucine for inhibiting the threonine dehydratase and L-valine forinhibiting metabolization of the ketobutyrate by the acetohydroxy acidsynthetase (Umbarger: Escherichia coli and Salmonella typhimurium, 1(1987) 352-367). A substance for inducing the threonine dehydratase geneand L-threonine as substrate for the dehydratase are preferably added tothe medium in addition. Preferably, IPTG(isopropyl-β-D-thiogalactopyranoside) is used as substance for inducingthe threonine dehydratase gene, so that an IPTG-induceable promoter isconnected in series with the altered threonine dehydratase gene in thevector. Such clones, which contain a deregulated threonine dehydratase,convert the L-threonine without inhibition into ketobutyrate. Since thefurther reaction of the ketobutyrate by acetohydroxy acid synthase isinhibited by the L-valine that has been added, the ketobutyrateaccumulates. As a result, the clones, which contain a deregulatedthreonine dehydratase, do not grow as well because of the known toxicityof ketobutyrate (La Rossa and Schloss, J. Biol. Chem. 259 (1984)8753-8757). The poorer growth is expressed by the formation of smallercolonies, more translucent colonies or colonies with a jagged outline.These colonies can easily be "abgeimpft" (inoculated off) and thereforeclones with deregulated dehydratase can easily be isolated.

This method of isolating can, of course, also be used for thosetransformants, which contain a cloned threonine dehydratase gene, whichhas been modified by mutations other than base exchange, so that thecorresponding enzyme no longer is subject to feedback inhibition.

An inventive alternative for accomplishing said objective consiststherein that, in a gene of a threonine dehydratase from Corynebacteriumglutamicum present in vitro, the amino acid alanine in position 257 ofthe amino acid sequence of the enzyme is replaced by the amino acidglycine (Table 4) (SEQ ID NO:8) or the amino acid methionine in position199 of the amino acid sequence of the enzyme is replaced by the aminoacid valine (Table 5) (SEQ ID NO:10) by a base exchange outside of thegene region coding for the allosteric domains of the enzyme. Aftertransformation of the threonine dehydratase gene so mutated into a hostcell producing threonine or L-isoleucine, the host cell increasinglyforms L-isoleucine.

A further alternative for accomplishing the inventive objective consiststherein that, in a threonine dehydratase gene from Corynebacteriumglutamicum, the amino acid, histidine, in position 278 of the amino acidsequence of the enzyme is replaced by the amino acid arginine by baseexchange outside of the gene region coding for the allosteric domains ofthe enzyme and the amino acid leucine in position 351 of the amino acidsequence of the enzyme is replaced by the amino acid serine (Table 6)(SEQ ID NO:12) by base exchange within the gene region coding for theallosteric domain of the enzyme. After transformation of the threoninedehydratase gene so mutated into a host cell producing threonine orL-isoleucine, this host cell also increasingly forms L-isoleucine.

Preferably Corynebacterium glutamicum and, particularly, the depositedDSM strain 8890 is suitable as host cell for the transformation of amutated threonine dehydratase gene or as production strain. Astransformants, the strains deposited with the DSM under the numbers 8889and 8891, for example, are obtainable, which increasingly secreteL-isoleucine into the fermentation medium. It is also beneficial to useproduction strains or host cells for the transformation, which no longersynthesize the wild type threonine dehydratase, which is still regulatedby L-isoleucine, so that only deregulated threonine dehydratase stillcatalyzes the conversion of L-threonine in the cells.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the biosynthetic pathway for L-isoleucine.

FIG. 2 shows a map of the plasmid pBM20.

FIG. 3 shows a map of the plasmid pKW0.

EXAMPLE

1. Production of Mutated Enzymes, Which are no Longer Subject toFeedback Inhibition

1.1 Selection of the Desired Enzymes

The known plasmid pBM1/Exo8, which codes the regulated threoninedehydratase of the wild type of Corynebacterium glutamicum (Mockel, etal., J. Bacteriol. 174 (1992) 8065-8072), was isolated by known methods(Sambrook et al., Molecular Cloning, A Laboratory Handbook, 1989, ColdSpring Harbor Laboratory Press). It was subsequently digested with therestriction enzyme EcoR1 and the fragment, containing the threoninedehydratase gene and consisting of 1573 base pairs, was isolated. Thisfragment was ligated in the EcoR1 cut site of the known vector pUC19(Vieira and Messing, Gene 19 (1976) 259-268). By these means, theplasmid pBM20 was obtained (FIG. 2), in which the threonine dehydratasegene from C. glutamicum, which itself is not expressed in E. coli,(Cordes et al., Gene 112 (1992) 113-116), is present in the standardlaboratory strain of E. coli JM109 under the control of the lacZpromoter of the original vector pUC19, which can be induced byisopropyl-D-thiogalactopyranoside.

In order to introduce non-directional mutations in the allostericdomains of the threonine dehydratase, two DNA primers (SEQ ID NOS 13 &NO:15) were synthesized by a known method in a separate procedure:

5'-primer: CGCAGCTGACTTCCATGGGGCAAGA

3'-primer: CCAGTGCCAAGCTTGCATGC

The 5' primer is homologous with the Ncol cut site (underlined) in thethreonine dehydratase gene. The 3' primer is homologous with the HindIIIrecognition site (underlined) of the multiple cloning site of the outputvector pUC19.

With both primers, a polymerase chain reaction was now carried out(Tindall and Kunkel, Biochemistry 27 (1988) 6008-6013), in which new DNAfragments, which contain mutations, result from the nonspecific reactionof the Taq polymerase (Leung et al., Technique 1 (1989) 11-15). In afinal volume of 100 μL, the reaction formulation contained 20 μL of 5'primer (25 μg/mL), 20 μL of 3' primer (25 μg/mL), 1 ng of template(pBM20), 100 μM of ATP, 100 μM of dNTP mix, 10 μL Taq 10× buffer (100 mMof tris, 15 mM of MgCl₂, 500 mM of KCl, 1 mg/mL of gelatin, pH 8.3), 0.5U of Taq polymerase (5 U/μL) and 0.5 mM of MnCl₂. The reactionconditions were set as follows: a time delay file of 94° C. (3 minutes),a thermo cycle file of 94° C. (1 minute), 55° C. (2 minutes), 72° C. (3minutes), a soak file of 4° C., a segment extension of 10 seconds, thenumber of cycles being 30. After the reaction, the DNA fragments werepurified (Sambrook et al., Molecular Cloning, A Laboratory Handbook,1989, Cold Spring Harbor Laboratory Press), digested with Ncol andHindIII and ligated with pBM20, from which the corresponding 743 basepairs Ncol-HindIII fragment, which codes for the allosteric domains ofthe wild type threonine dehydratase, had previously been cut out.

With these in vitro produced ligation products, which also containmutated threonine dehydratase genes, E. coli JM109 was now transformedby known methods (Hanahan, Techniques for Transformation of E. coli(1985) DNA Cloning, volume 1, pp. 109-136, IRL Press Oxford) and about10,000 clones, containing plasmid, were obtained on Luria-Bertani medium(Lennox, Virology 1 (1955) 190-206), which contained 50 μg/mL ofampicillin.

In order to identify the clones, which code for a threonine dehydratase,which can no longer be inhibited by feedback, a Luria-Bertani medium wasprepared, which additionally contained 40 mM of L-threonine as substratefor the threonine dehydratase, as well as 1 mM ofisopropyl-β-D-thiogalactopyranoside for inducing the lacZ-inducedexpression of the threonine dehydratase gene. As is well known, theLuria-Bertani medium already contains sufficient L-valine for inhibitingthe activity of the acetohydroxy acid synthase in E. coli (Umbarger,Biosynthesis of Branched-Chain Amino Acids (1987) Escherichia coli andSalmonella typhimurium, vol. 1, pp. 352-367, American Society forMicrobiology, Washington, DC) in order to attain the highest possibleaccumulation of α-ketobutyrate by the method described here. Aside fromthe Luria-Bertani medium, which contained threonine andisopropyl-β-D-thiogalactopyranoside, the same medium was prepared as acontrol without isopropyl-β-D-thiogalactopyranoside, so that there wasno induction of the threonine dehydratase. Subsequently, the clones ofE. coli JM109, which contained the pBM20 derivatives to be tested, werestamped on these plates in the conventional manner and incubated for 24hours at 37° C. After that, it was possible to identify on the medium,which contained isopropyl-β-D-thiogalactopyranoside 60 abnormallygrowing colonies, which were distinguished by a lesser growth or palercolonies or colonies having a jagged edge, but grew normally on thecontrol medium without isopropyl-β-D-thiogalactopyranoside. These cloneswere subjected individually to an identical follow-up test and,subsequently, 6 of the clones with a pronounced growth retardation weresubjected to a biochemical test for characterizing the regulation of thethreonine dehydratases containing them.

1.2 Characterizing the Inhibition of the Enzymes

Six of the recombinant clones of E. coli JM109, so obtained, wereinoculated in 100 mL of LB liquid medium, which additionally contained50 μg of ampicillin/mL and incubated at 37° C. The optical density(OD₆₀₀ nm) was followed. When the OD reached a value of 0.5, theisopropyl-β-D-thiogalactopyranoside was added until it reached a finalconcentration of 0.5 mM. After incubating for a further hour at 37° C.,the cells were harvested by centrifuging, washed once with a pH 7.0buffer (0.1 M potassium phosphate, 0.5 mM L-isoleucine, 0.2 mM pyridoxalphosphate) and taken up in 1 mL of the same buffer and disintegrated byan ultrasonic treatment in the Branson-Sonifier W250 (3 minutes, pulsedwith an interval of 20% and an output of 2). For separating the cellfragments, the homogenate was centrifuged for 10 minutes at 13,000 rpmand 4° C. in a Sigma refrigerated centrifuge. Subsequently, theresulting clear supernatant (crude extract) was used in the enzyme testfor determining threonine dehydratase activity and regulation.

In a final volume of 0.8 mL, the enzyme test contained 0.1 M potassiumphosphate (pH 8.2), 1 mM of pyridoxal phosphate, 40 mM of L-threonineand crude extract. The mixture was incubated at 30° C. and 200 μLsamples were taken after 0 and 30 minutes. The threonine dehydratasereaction was terminated in each case by the addition of 1 mL of reagent.This consisted of 1 g of semicarbazide plus 0.9 g of sodium acetate in100 mL of water. After incubating for 15 minutes at 30° C., 3 mL ofwater were added and the extinction was determined at 254 nm in a ZeissPM6 spectrophotometer. Controls and calibration values with 0 to 1.5μmoles of ketobutyrate were treated identically and the amount ofketobutyrate, actually formed in the enzyme test by the threoninedehydratase, was determined from a calibration curve. Parallel to this,identical formulations were carried out, which contained 5 mM ofL-isoleucine for testing for the inhibition of threonine dehydratase.Independently of this determination, the protein content of the crudeextracts was determined as described (Bradford, Anal. Biochem. 72 (1976)248-254). The specific activities obtained and the degree of inhibitionare evident from Table 7.

                  TABLE 7                                                         ______________________________________                                        Characterization of Mutated Threonine Dehydratases and the                     Ability to Inhibit Them by L-Isoleucine                                                      Specific Activity of                                                                           Threonine Dehydratase                          (μmoles/min/mg of Protein)                                               Clone           -Ile   +5 mM Ile                                              ______________________________________                                        wild type       1.561  0.240                                                    28 1.771 1.907                                                                16 0.412 0.251                                                                31 1.932 0.590                                                                50 0.220 0.132                                                                54 0.28 0.136                                                                 14 1.708 1.989                                                              ______________________________________                                    

It can be seen directly that the mutated enzymes 38 and 14 are notinhibited allosterically by L-isoleucine. In the presence ofL-isoleucine, the enzyme of mutant 31 still has 30% of the activity,whereas the enzyme of the wild type has only 15% of the activity underthese conditions.

1.3 Characterization of the Mutation Site of the Enzyme

To determine the mutations in the enzymes produced, the plasmids, codingfor threonine dehydratase, were isolated from E. coli JM109 clones bystandard methods. The sequencing of the mutated region of the threoninedehydratase gene was carried out with the help of the didesoxynucleotidetermination method of Sanger et al. (Sanger et al., Proceedings of theNational Academy of Sciences, U.S.A. (1977) 5463-5467).Fluorescein-labeled sequence-specific nucleotides were synthesized asprimers following the directions of Pharmacia (Pharmacia, Uppsala,Sweden). These primers were purified by means of high-pressure liquidchromatography with a gradient of 5-30% acetonitrile in 100 mM oftriethylamine acetate of pH 7.0 and 5 μm of the Pharmacia SuperPac®Pep-S. The primers were used in a standard sequencing reaction and,during the electrophoresis, the reaction products were identified byfluorescence detection on the A. L. F. sequencer (Pharmacia, Uppsala,Sweden) and detected automatically and the resulting sequence wasrecorded.

Primers Used for the Sequencing

The position of the bases refers to the sequence in FIG. 3 in Mockel etal., J. Bacteriol. 174 (1992) 8065-8072.

Base 957-958(SEQ ID NO:15): 5'-Fluorescein-d(GGTCAGGGCACCGTGGCTGCTG)-3'

Base 1172-1191 (SEQ ID NO:16): 5'-Fluorescein-d(GGAGACTGTTGATCCCTTTG)-3'

Base 1381-1400 (SEQ ID NO:17): 5'-Fluorescein-d(CCTTTGCACCTGGTTCTGTC)-3'

Base 1586-1607 (SEQ ID NO:18):5'-Fluorescein-d(CCTCAAGCGCAACAACCGTGAG)-3'

The sequence of the individual threonine dehydratase genes, determinedthis manner, was compared with the known sequence of the wild type(Mockel et J. Bacteriol. 174 (1992) 8065-8072). The individual baseexchanges of the biochemically characterized threonine dehydratase genesare given in Table 8. The altered codons were translated into thecorresponding amino acids by means of the universal genetic code and theamino acid exchanges, which were thus detected and lead to alteredregulation of the threonine dehydratase, are also listed in Table 8.Mutations with a deregulated phenotype were obtained over the whole ofthe region used for the mutation. In mutant 14, there is a doublemutation, which shows that several exchanges of amino acids in oneenzyme can also lead to the desired deregulated phenotype.

                  TABLE 8                                                         ______________________________________                                        Genetic Characterization of Threonine Dehydratase with Altered                  Allosteric Regulation                                                              Nucleotide Exchange                                                                            Amino Acid Exchange                                   Mutants                                                                              Position                                                                              Wild Type                                                                              Mutants                                                                             Position                                                                            Wild Type                                                                            Mutants                            ______________________________________                                        38     1398    GTC      GCC   323   Val    Ala                                  16 1559 GAT GGT 377 Asp Gly                                                   31 1579 TTT TGT 383 Phe Cys                                                   50 1200 GCA GGA 257 Ala Gly                                                   54 1026 ATG GTG 199 Met Val                                                   14 1264 CAC CGC 278 His Arg                                                    1483 TTG TCG 351 Leu Ser                                                   ______________________________________                                    

2. Determination of the Isoleucine Secretion by Deregulated Enzymes

The alleles obtained were recloned in E. coli/C. glutamicum shuttlevectors, in order to be able to express them in this manner in athreonine producer of C. glutamicum.

For this purpose, the E. coli/C. glutamicum shuttle vector pKWO wasproduced to begin with by known cloning methods (Sambrook et al.,Molecular Cloning, A Laboratory Handbook, 1989, Cold Spring HarborLaboratory Press). The resulting vector of 9.5 kb is shown in FIG. 1. Itcontains (i) the C. glutamicum replicon pGA2 (Sonnen,Molekulargenetische Charakterisierung von Phagen-Wirt-Beziehungen beicoryneformen Aminosaure-Produzenten (Molecular Genetic Characterizationof Phage-Host Relationships in the Case of Coryneform Amino AcidProducers), dissertation, Technical University of Darmstadt, 1991),which leads to only a low number of copies in C. glutamicum, (ii) the E.coli replicon from pOU71 (Larsen et al., Gene 28 (1984) 45-54), whichleads to one copy per cell at 30° C. and up to 1000 copies per cell at42° C., (iii) the tetracycline resistance from pHY163 (Ishiwa andShibahara, Jpn. J. Genet 60 (1985) 485-498) and (iv) the cos site frompBTI-1 (Boehringer, Mannheim). The ilvA alleles 14, 16 and 18, as wellas the wild type allele, were isolated as EcoRI fragments from pBM20derivatives, produced under 1.1, and ligated with the EcoRI-restrungvector pKWO, in order to result in pKWOilvA, pKWOilvA14, pKWOilvA16,pKWOilvA38. With these plasmids, the threonine producerMH20-22B::pSUR5-DR1 (Reinscheid et al., Appl. Env. Microbiol. (1994), inpress) was transformed by means of electroporation (Liebl et al., FEMSMicrobiol. Lett. 65 (1985) 299-304). MH20-22B has been filed with theDeutschen Sammlung fir Mikroorganismen (German Collection forMicroorganisms) under the number DSM 6870, as were MH20-22B::pSUR5-DR1(DSM 8890) and MH20-22B::pSUR5-DR1 pKWOilvA16 (DSM 8889). These strainswere cultured in the minimum medium CGXII as described (Keilhauer etal., J. Bacteriol. 175 (1993) 5595-5603), and the amino acids, which hadaccumulated in the culture medium after a 72-hour incubation, weredetermined. Table 9 clearly shows that the formation of L-isoleucine isincreased by the mutant alleles.

                  TABLE 9                                                         ______________________________________                                        Effect of the Expression of the Alleles of the Mutants 38, 14 and 16 on        the Production of Amino Acid with G. glutamicum                                                                Lys                                                                                 Strain Thr (mM) Ile                   ______________________________________                                        MH20-22B              0       76      1                                         MH20-22B::pSUR5-DR1 36 26 16                                                  MH20-22B::pSUR5-DR1 pKWOilvA 14 26 40                                         MH20-22B::pSUR5-DR1 pKWOiivA14  0 24 55                                       MH20-22B::pSUR5-DR1 pKWOilvA16  0 22 50                                       MH20-22B::pSUR5-DR1 pKWOilvA38  0 20 53                                     ______________________________________                                    

In a further experiment, the wild type allele and the allele of mutant38 were incorporated in the high copy pendulum vector pECM3 (A. Schafer,Diploma Thesis, 1991, University of Bielefeld). The fragments once againwere isolated as EcoRI fragments from the pBM20 derivatives producedunder 1.1 and ligated with the EcoRI-restrung vector, in order to resultin pECM3ilvA and pECM3ilvA38. With these plasmids, the threonineproducer MH20-22B::pSUR5-DR1 (Reinscheid et al., Appl. Env. Microbiol.(1994), in press) was transformed by means of electroporation (Liebl etal., FEMS Microbiol. Lett. 65 (1985) 299-304). MH20-22B has been filedwith the Deutschen Sammlung fur Mikroorganismen (German Collection forMicroorganisms) under the number DSM 6870, as were MH20-22B::pSUR5-DR1(DSM 8890) and MH20-22B::pSUR5-DR1 pECM3ilvA38 (DSM 8891). These strainswere cultured in the MH20-22B minimum medium, as described (Schrumpf etal. Appl. Microbiol. Biotechnol. 37 (1992) 566-571) and the amino acids,which had accumulated in the culture medium after a 72-hour incubation,were determined. Table 10 once again clearly shows that the formation ofL-isoleucine is increased by the respective mutant allele.

                  TABLE 10                                                        ______________________________________                                        Effect of the Expression of the Alleles of Mutant 38 on the                     Production of Amino Acid with C. glutamicum                                                                  Lys                                            Strain Thr (mM) Ile                                                         ______________________________________                                        MH20-22B            0        158     2                                          MH20-22B::pSUR5-DR1 53  61 29                                                 MH20-22B::pSUR5-DR1 pECM3i1vA 0 55 88                                         MH20-22B::pSUR5-DR1 pECM3i1vA38 0 59 125                                    ______________________________________                                    

                                      TABLE 1                                     __________________________________________________________________________    Mutant 38 (SEQ ID NO:1)                                                       __________________________________________________________________________             .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                             -          .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                        -          .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                             -          .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                        -          .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                             -          .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                        -          .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                             -          .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG  400                        -          .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  I  T  Y  V                               -          .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCGTGCC  500                       S  I  N  S  P  G  V  M  A  S  G  A  I  I  I  P  A    23                        -          .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  Q  I  A  Q  A  R  I  S  S  V  I  A  P  T                             -          .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  I  S  I  F  I  G  A  I  I     56                        -          .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  L  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                              -          .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                        A  L  N  S  G  A  Q  S  P  V  E  Q  R  D  A  C  I   90                        -          .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGYGTGCA                              V  A  A  S  A  G  N  H  A  Q  G  V  A  Y  V  C                               -          .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  R  I  Y  V  P  V  Q  T  P  N   123                        -          .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                             -          .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATG  900                         V  T  G  N  N  F  D  E  A  S  A  A  A  N  E  D    156                        -          .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                              -          .         .        BglII      .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                        -          .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGATGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  H  V  M  V  P  V  G  G  G  G                               -          .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  I  A   223                        -          .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  M  Q  A  A  L  H                             -          .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                        -          .         BglII     .         .         .                         CAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            A  E  V  K  R  V  G  D  L  N  Y  T  I  V  E  K  N                              -          .         .         .         .         .                         CAGGGTCGCGTGCACATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  H  M  M  S  A  T  E  G  A  V  C  T  C  290                        -          .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGGCG                              M  L  D  L  Y  Q  N  E  G  I  I  A  E  P  A  G                               -          .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGCC 1400                       A  L  S  I  A  G  L  K  E  M  S  F  A  P  G  S  A    323                                                         ---------------                             -          .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  D  V  L  R  Y  A  E                             -          .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTTGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  L  K  H  Y  I  L    356                        -          .         .         .         .    EcoRV.                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  N  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                              -          .         .         .         .         .                         CTGGGACCGGATGATGACATCACGCTGTTTGAGTACCTCAAGCGCAACAA 1600                       L  G  P  D  D  D  I  T  L  F  E  Y  L  K  R  H  N   390                        -          .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTGAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                               -          .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  R   423                        -          .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  E  P  G  T  P  E  Y  E  Y  L  T  *                                      -          .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                         436                                                                          -          .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                             -          .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                        -          .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                   __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________    Mutant 16 (SEQ ID NO:3)                                                       __________________________________________________________________________             .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                             -          .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                        -          .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                             -          .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                        -          .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                             -          .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                        -          .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                             -          .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG  400                        -          .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  E  T  Y  V                               -          .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCGTGCC  500                       S  E  K  S  P  G  V  M  A  S  G  A  I  I  I  R  A    23                        -          .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  Q  I  A  Q  A  R  I  S  S  V  I  A  P  T                             -          .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  L  S  C  E  T  G  A  E  I     56                        -          .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  L  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                              -          .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                        A  L  N  S  G  A  Q  S  P  Q  E  Q  P  Q  A  G  I   90                        -          .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGTGTGCA                              V  A  A  S  A  G  N  H  A  Q  G  V  A  Y  V  C                               -          .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  R  I  Y  V  P  V  Q  T  P  K   123                        -          .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                             -          .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATG  900                         V  T  G  N  N  F  D  E  A  S  A  A  A  H  E  D    156                        -          .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                              -          .         .        BglII      .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                        -          .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGATGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  H  V  M  V  P  V  G  G  G  G                               -          .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  T  A   223                        -          .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  M  Q  A  A  L  H                             -          .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                        -          .         BglII     .         .         .                         CAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            A  E  V  K  R  V  G  D  L  N  Y  T  I  V  E  K  N                              -          .         .         .         .         .                         CAGGGTCGCGTGCACATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  H  M  M  S  A  T  E  G  A  V  C  T  E  290                        -          .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGGCG                              M  L  D  L  Y  Q  N  E  G  I  I  A  E  P  A  G                               -          .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGTC 1400                       A  L  S  I  A  G  L  K  E  M  S  F  A  P  G  S  V  323                                                           ---------------                             -          .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  D  V  L  R  Y  A  E                             -          .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTTGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  L  K  H  Y  F  L    356                        -            .         .         .         .    EcoR                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  N  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                              - V.         .         .         .         .                                 CTGGGACCGGGTGATGACATCACGCTGTTTGAGTACCTCAAGCGCAACAA 1600                        L  G  P  G  D  D  I  T  L  F  E  Y  L  K  R  H  N   390                       -          .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTGAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                               -          .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  R   423                        -          .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  E  P  G  T  P  E  Y  E  Y  L  T  *                                      -          .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                         436                                                                          -          .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                             -          .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                        -          .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                   __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Mutant 31 (SEQ ID NO:5)                                                       __________________________________________________________________________             .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                             -          .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                        -          .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                             -          .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                        -          .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                             -          .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                        -          .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                             -          .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG   400                       -          .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  E  T  Y  V                               -          .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCGTGCC  500                       S  E  K  S  P  G  V  M  A  S  G  A  E  I  I  R  A    23                        -          .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  Q  T  A  Q  A  R  I  S  S  V  I  A  P  T                             -          .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  L  S  C  E  T  G  A  E  I     56                        -          .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  L  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                              -          .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                       A  L  N  S  G  A  Q  S  P  Q  E  Q  R  D  A  G  I    90                        -          .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGTGTGCA                              V  A  A  S  A  G  N  H  A  Q  G  V  A  Y  V  C                               -          .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  R  I  Y  V  P  V  Q  T  P  K   123                        -          .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                             -          .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATG  900                         V  T  G  N  N  F  D  E  A  S  A  A  A  H  E  D    156                        -          .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                              -          .         .        BglII      .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                        -          .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGATGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  N  V  M  V  P  V  G  G  G  G                               -          .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  T  A   223                        -          .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  H  Q  A  A  L  H                             -          .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                        -          .         BglII     .         .         .                         CAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            A  E  V  K  R  V  G  D  L  N  Y  T  I  V  E  K  N                              -          .         .         .         .         .                         CAGGGTCGCGTGCACATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  H  M  M  S  A  T  E  G  A  V  C  T  E  290                        -          .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGCCG                              M  L  D  L  Y  Q  R  E  G  I  I  A  E  P  A  G                               -          .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGTC 1400 - A  L  S  I  A                                      G  L  K  E  M  S  F  A  P  G  S  V   323                                          --------------                              -          .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  Q  V  L  R  Y  A  E                             -          .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTTGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  L  K  H  Y  F  L    356                        -          .         .         .         .    EcoRV.                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  N  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                              -          .         .         .         .         .                         CTGGGACCGGATGATGACATCACGCTGTGTGAGTACCTCAAGCGCAACAA 1600                        L  G  P  D  D  D  I  T  L  C  E  Y  L  K  R  N  N   390                       -          .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTGAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                               -          .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  A   423                        -          .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  E  P  G  T  P  C  Y  E  Y  L  T  *                                      -          .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                         436                                                                          -          .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                             -          .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                        -          .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                   __________________________________________________________________________

                                      TABLE 4                                     __________________________________________________________________________    Mutant 50 (SEQ ID NO:4)                                                       __________________________________________________________________________              .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                            -           .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                       -           .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                            -           .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                       -           .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                            -           .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                       -           .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                            -           .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG  400                       -           .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  I  T  Y  V                              -           .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCGTGCC  500                       S  E  K  S  P  G  V  M  A  S  G  A  E  I  I  H  A    23                       -           .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  G  I  A  Q  A  R  I  S  S  V  I  A  P  T                            -           .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  L  S  I  I  T  G  A  I  I     56                       -           .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  L  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                             -           .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                        A  L  N  S  G  A  Q  S  P  Q  I  Q  F  D  A  G  I   90                       -           .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGTGTGCA                              V  A  A  S  A  G  H  H  A  Q  G  V  A  Y  V  C                              -           .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  R  I  Y  V  P  V  Q  T  P  K   123                       -           .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                            -           .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATG  900                         V  T  G  N  N  F  D  E  A  S  A  A  A  H  E  D    156                       -           .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                             -           .         .         .         .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                       -           .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGATGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  H  V  M  V  P  V  G  G  G  G                              -           .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  T  A   223                       -           .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  M  Q  A  A  L  H                            -           .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                       -           .         BglII     .         .         .                         GAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            G  E  V  K  R  V  G  D  L  N  Y  T  I  V  E  K  N                             -           .         .         .         .         .                         CAGGGTCGCGTGCACATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  H  M  M  S  A  T  C  G  A  V  C  T  E  290                       -           .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGGCG                              M  L  D  L  Y  Q  N  E  G  I  I  A  E  P  A  G                              -           .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGTC 1400                       A  L  S  I  A  G  L  K  E  M  S  F  A  P  G  S  V   323                                                          --------------                             -           .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  D  V  L  R  Y  A  E                            --------------------------------------------------                            -           .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTTGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  L  K  H  Y  F  L    356                       --------------------------------------------------                            -           .         .         .         .    EcoRV.                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  N  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                             --------------------------------------------------                            -           .         .         .         .         .                         CTGGGACCGGATGATGACATCACGCTGTTTGAGTACCTCAAGCGCAACAA 1600                        L  G  P  D  D  D  I  T  L  F  E  Y  L  K  R  N  H  390                       --------------------------------------------------                            -           .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTCAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                              --------------------------------------------------                            -           .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  R   423                       --------------------------------------------------                            -           .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  C  P  G  T  P  E  Y  C  Y  L  T  *                                     ---------------------------------------                                       -           .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                       436                                                                           -           .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                            -           .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                       -           .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                  __________________________________________________________________________

                                      TABLE 5                                     __________________________________________________________________________    Mutant 54 (SEQ ID NO:10)                                                      __________________________________________________________________________             .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                             -          .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                        -          .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                             -          .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                        -          .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                             -          .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                        -          .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                             -          .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG  400                        -          .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  E  T  Y  V                               -          .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCGTGCC  500                       S  F  F  S  P  G  V  M  A  S  G  A  I  I  I  R  A    23                        -          .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  Q  I  A  Q  A  R  I  S  S  V  I  A  P  T                             -          .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  I  S  I  I  T  G  A  I  I     56                        -          .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  I  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                              -          .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                        A  L  N  S  G  A  Q  S  P  Q  T  Q  K  D  A  G  I   90                        -          .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGTGTGCA                              V  A  A  S  A  G  N  H  A  Q  G  V  A  Y  V  C                               -          .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  T  I  Y  V  P  V  Q  T  P  K   123                        -          .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                             -          .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATG  900                         V  T  G  H  N  F  D  E  A  S  A  A  A  H  E  D    156                        -          .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                              -          .         .        BglII      .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                        -          .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGGTGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  H  V  V  V  P  V  G  G  G  G                               -          .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  T  A   223                        -          .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  M  Q  A  A  L  H                             -          .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                        -          .         BglII     .         .         .                         CAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            A  E  V  K  R  V  G  D  L  H  Y  T  I  V  E  K  N                              -          .         .         .         .         .                         CAGGGTCGCGTGCACATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  H  M  M  S  A  T  E  G  A  V  C  T  E   290                       -          .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGGCG                              M  L  D  L  Y  Q  N  E  G  I  I  A  E  P  A  G                               -          .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGTC 1400                       A  L  S  I  A  G  L  K  E  M  S  F  A  P  G  S  V   323                                                          ---------------                             -          .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  D  V  L  R  Y  A  E                             -          .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTTGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  L  K  H  Y  F  L    356                        -          .         .         .         .    EcoRV.                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  H  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                              -          .         .         .         .         .                         CTGGGACCGGATGATGACATCACGCTGTTTGAGTACCTCAAGCGCAACAA 1600                        L  G  P  D  D  D  I  I  L  F  E  Y  L  K  R  N  N  390                        -          .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTGAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                               -          .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  R   423                        -          .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  E  P  G  T  P  C  Y  E  Y  L  T  *                                      -          .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                        436                                                                           -          .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                             -          .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                        -          .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                   __________________________________________________________________________

                                      TABLE 6                                     __________________________________________________________________________    Mutant 14 (SEQ ID NO:12)                                                      __________________________________________________________________________             .         .         .         .         .                              CGCCATTGCTGAGCATTGAGCTGCCTTCAGAGCTGCCTGGCCAGGTTTCG                             -          .         .         .         .         .                         TTTCCATCGACTGGATTTCCATCATCATCAAGGATCTGTGATGAGGTGAT  100                        -          .         .         .         .         .                         GTTGTCTGAGAGCTGTGTCAGTGCGTCAGAGGACTGAGCCTGGGCAACTG                             -          .         .         .         .         .                         GAGTGAACACGGACAATGCCACAGCGCTTGCTGTAACAAGGGTCAAAGTA  200                        -          .         .         .         .         .                         CTTCGACGCAAAGACAAAACTTTTCTCCTGGCAATAAATATGCGGATTTA                             -          .         .         .         .         .                         CTATGGAAACAAGATAGAAGATTGGATAGCGAAAGCTATCCTCAACTCGT  300                        -          .         .         .         .         .                         GGAAAGTGTAGTGCCACAACCACAGTATTGGCTAGAAAACAATCTATAGC                             -          .         .         .         .         .                         ATTGTTCTACAAAGAGCTTGTTGGAAATAAAACCTATGCCAAAGTAGGTG  400                        -          .         .    HindII         .         .                         CAATTCTAGGAGAAGATTACACTAGTCAACCATGAGTGAAACATACGTGT                                                            M  S  E  T  Y  V                               -          .         .         .         .         .                         CTGAGAAAAGTCCAGGAGTGATGGCTAGCGGAGCGGAGCTGATTCCTGCC  500                       S  E  K  S  P  G  V  M  A  S  G  A  E  I  I  R  A    23                        -          .         .         .         .         .                         GCCGACATTCAAACGGCGCAGGCACGAATTTCCTCCGTCATTGCACCAAC                             A  D  I  Q  T  A  Q  A  R  I  S  S  V  I  A  P  T                             -          .         .         .         .         .                         TCCATTGCAGTATTGCCCTCGTCTTTCTGAGGAAACCGGAGCGGAAATCT  600                         P  L  Q  Y  C  P  R  I  S  I  E  T  G  A  E  I     56                        -          .         .         .         .         .                         ACCTTAAGCGTGAGGATCTGCAGGATGTTCGTTCCTACAAGATCCGCGGT                            Y  L  K  R  E  D  L  Q  D  V  R  S  Y  K  I  R  G                              -          .         .         .         .         .                         GCGCTGAACTCTGGAGCGCAGTCACCCCAAGAGCAGCGCGATGCAGGTAT  700                        A  L  N  S  G  A  Q  S  P  Q  E  Q  R  D  A  G  I   90                        -          .      BstEII       .         .         .                         CGTTGCCGCATCTGCAGGTAACCATGCCCAGGGCGTGGCCTATGTGTGCA                              V  A  A  S  A  G  N  H  A  Q  G  V  A  Y  V  C                               -          .         .         .         .         .                         AGTCCTTGGGCGTTCAGGGACGCATCTATGTTCCTGTGCAGACTCCAAAG  800                       K  S  L  G  V  Q  G  R  I  Y  V  P  V  Q  T  P  K   123                        -          .         .         .         .         .                         CAAAAGCGTGACCGCATCATGGTTCACGGCGGAGAGTTTGTCTCCTTGGT                             Q  K  R  D  R  I  M  V  H  G  G  E  F  V  S  L  V                             -          .         .         .         .         .                         GGTCACTGGCAATAACTTCGACGAAGCATCGGCTGCAGCGCATGAAGATC  900                         V  T  G  N  N  F  D  E  A  S  A  A  A  H  E  D    156                        -          .         .         .         .         .                         CAGAGCGCACCGGCGCAACGCTGATCGAGCCTTTCGATGCTCGCAACACC                            A  E  R  T  G  A  T  L  I  E  P  F  D  A  R  N  T                              -          .         .        BglII      .         .                         GTCATCGGTCAGGGCACCGTGGCTGCTGAGATCTTGTCGCAGCTGACTTC 1000                        V  I  G  Q  G  T  V  A  A  E  I  L  S  Q  L  T  S  190                        -          .         .         .         .         .                         CATGGGCAAGAGTGCAGATCACGTGATGGTTCCAGTCGGCGGTGGCGGAC                              M  G  K  S  A  D  H  V  M  V  P  V  G  G  G  G                               -          .         .         .         .         .                         TTCTTGCAGGTGTGGTCAGCTACATGGCTGATATGGCACCTCGCACTGCG 1100                       L  L  A  G  V  V  S  Y  M  A  D  M  A  P  R  T  A   223                        -          .         .         .         .         .                         ATCGTTGGTATCGAACCAGCGGGAGCAGCATCCATGCAGGCTGCATTGCA                             I  V  G  I  E  P  A  G  A  A  S  M  Q  A  A  L  H                             -          .         .         .         .         .                         CAATGGTGGACCAATCACTTTGGAGACTGTTGATCCCTTTGTGGACGGCG 1200                         N  G  G  P  I  T  L  E  T  V  D  P  F  V  D  G    256                        -          .         BglII     .         .         .                         CAGAGGTCAAACGTGTCGGAGATCTCAACTACACCATCGTGGAGAAGAAC                            A  E  V  K  R  V  G  D  L  N  Y  T  I  V  E  K  N                              -          .         .         .         .         .                         CAGGGTCGCGTGCGCATGATGAGCGCGACCGAGGGCGCTGTGTGTACTGA 1300                        Q  G  R  V  R  M  M  S  A  T  E  G  A  V  C  I  E  290                        -          .         .         .         .         .                         GATGCTCGATCTTTACCAAAACGAAGGCATCATCGCGGAGCCTGCTGGCG                              M  L  D  L  Y  Q  N  E  G  I  I  A  E  P  A  G                               -          .         .         .         .         .                         CGCTGTCTATCGCTGGGTTGAAGGAAATGTCCTTTGCACCTGGTTCTGTC 1400                       A  L  S  I  A  G  L  K  E  M  S  F  A  P  G  S  V   323                                                          ---------------                             -          .         .         .         .         .                         GTGGTGTGCATCATCTCTGGTGGCAACAACGATGTGCTGCGTTATGCGGA                             V  V  C  I  I  S  G  G  N  N  D  V  L  R  Y  A  E                             -          .         .         .         .         .                         AATCGCTGAGCGCTCCTTGGTGCACCGCGGTTCGAAGCACTACTTCTTGG 1500                         I  A  E  R  S  L  V  H  R  G  S  K  N  Y  F  L    356                        -          .         .         .         .    EcoRV.                         TGAACTTCCCGCAAAAGCCTGGTCAGTTGCGTCACTTCCTGGAAGATATC                            V  N  F  P  Q  K  P  G  Q  L  R  H  F  L  E  D  I                              -          .         .         .         .         .                         CTGGGACCGGATGATGACATCACGCTGTTTGAGTACCTCAAGCGCAACAA 1600                        L  G  P  D  D  D  I  T  L  F  E  Y  L  K  R  N  N  390                        -          .         .         .         .         .                         CCGTGAGACCGGTACTGCGTTGGTGGGTATTCACTTGAGTGAAGCATCAG                              R  E  T  G  T  A  L  V  G  I  H  L  S  E  A  S                               -          .         .         .         .         .                         GATTGGATTCTTTGCTGGAACGTATGGAGGAATCGGCAATTGATTCCCGT 1700                       G  L  D  S  L  L  E  R  M  E  E  S  A  I  D  S  R   423                        -          .         .         .         .         .                         CGCCTCGAGCCGGGCACTCCTGAGTACGAATACTTGACCTAAACATAGCT                             R  L  E  P  G  T  P  E  Y  E  Y  L  T  *                                      -          .         .         .         .         .                         GAAGGCCACCTCAATCGAGGTGGCCTTTTTCTAGTTTCGGGTCAGGATCG 1800                        436                                                                           -          .         .         .         .         .                         CAAAGCCCCACGGCTGAAGGGTTGTGGAGGTGTCGGTGACGGTGGGGGAA                             -          .         .         .         .         .                         GTGAAGCTGTAAATCAGCTCGCCGCCAAGCGGGACGGTGATGGTGTCGTC 1900                        -          .    EcoRI     .                                                  GGAGAAATTCGCCAGAATTCGGCCG                                                   __________________________________________________________________________

    __________________________________________________________________________    #             SEQUENCE LISTING                                                   - -  - - (1) GENERAL INFORMATION:                                             - -    (iii) NUMBER OF SEQUENCES: 18                                          - -  - - (2) INFORMATION FOR SEQ ID NO:1:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -         (xi) SEQUENCE DESCRIPTION: SEQ - #ID NO:1:                        - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -            #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG ATG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Met Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GCA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Ala Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CAC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val His Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GCC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Ala Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TTG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Leu Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GAT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Asp Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TTT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Phe Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:2:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                               - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Met Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Ala Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val His Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Ala Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Leu Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Asp Asp As - #p Ile Thr Leu Phe Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:3:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                               - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -             #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG ATG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Met Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GCA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Ala Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CAC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val His Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GTC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Val Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TTG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Leu Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GGT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Gly Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TTT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Phe Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:4:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                               - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Met Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Ala Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val His Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Val Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Leu Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Gly Asp As - #p Ile Thr Leu Phe Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:5:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                               - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -             #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG ATG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Met Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GCA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Ala Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CAC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val His Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GTC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Val Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TTG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Leu Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GAT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Asp Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TGT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Cys Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:6:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                               - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Met Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Ala Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val His Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Val Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Leu Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Asp Asp As - #p Ile Thr Leu Cys Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:7:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                               - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -             #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG ATG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Met Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GGA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Gly Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CAC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val His Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GTC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Val Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TTG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Leu Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GAT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Asp Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TTT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Phe Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:8:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                               - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Met Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Gly Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val His Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Val Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Leu Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Asp Asp As - #p Ile Thr Leu Phe Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:9:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                               - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -             #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG ATG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Met Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GCA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Ala Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CAC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val His Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GTC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Val Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TTG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Leu Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GAT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Asp Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TTT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Phe Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:10:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                              - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Met Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Ala Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val His Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Val Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Leu Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Asp Asp As - #p Ile Thr Leu Phe Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:11:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1925 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 432..1739                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                              - - CGCCATTGCT GAGCATTGAG CTGCCTTCAG AGCTGCCTGG CCAGGTTTCG TT -             #TCCATCGA     60                                                                 - - CTGGATTTCC ATCATCATCA AGGATCTGTG ATGAGGTGAT GTTGTCTGAG AG -            #CTGTGTCA    120                                                                 - - GTGCGTCAGA GGACTGAGCC TGGGCAACTG GAGTGAACAC GGACAATGCC AC -            #AGCGCTTG    180                                                                 - - CTGTAACAAG GGTCAAAGTA CTTCGACGCA AAGACAAAAC TTTTCTCCTG GC -            #AATAAATA    240                                                                 - - TGCGGATTTA CTATGGAAAC AAGATAGAAG ATTGGATAGC GAAAGCTATC CT -            #CAACTCGT    300                                                                 - - GGAAAGTGTA GTGCCACAAC CACAGTATTG GCTAGAAAAC AATCTATAGC AT -            #TGTTCTAC    360                                                                 - - AAAGAGCTTG TTGGAAATAA AACCTATGCC AAAGTAGGTG CAATTCTAGG AG -            #AAGATTAC    420                                                                 - - ACTAGTCAAC C ATG AGT GAA ACA TAC GTG TCT GAG - #AAA AGT CCA GGA        GTG     470                                                                                  Met Ser Glu - #Thr Tyr Val Ser Glu Lys Ser Pro Gly Val                         1   - #            5      - #            10                     - - ATG GCT AGC GGA GCG GAG CTG ATT CGT GCC GC - #C GAC ATT CAA ACG GCG          518                                                                       Met Ala Ser Gly Ala Glu Leu Ile Arg Ala Al - #a Asp Ile Gln Thr Ala                15             - #     20             - #     25                          - - CAG GCA CGA ATT TCC TCC GTC ATT GCA CCA AC - #T CCA TTG CAG TAT TGC          566                                                                       Gln Ala Arg Ile Ser Ser Val Ile Ala Pro Th - #r Pro Leu Gln Tyr Cys            30                 - # 35                 - # 40                 - # 45       - - CCT CGT CTT TCT GAG GAA ACC GGA GCG GAA AT - #C TAC CTT AAG CGT GAG          614                                                                       Pro Arg Leu Ser Glu Glu Thr Gly Ala Glu Il - #e Tyr Leu Lys Arg Glu                            50 - #                 55 - #                 60              - - GAT CTG CAG GAT GTT CGT TCC TAC AAG ATC CG - #C GGT GCG CTG AAC TCT          662                                                                       Asp Leu Gln Asp Val Arg Ser Tyr Lys Ile Ar - #g Gly Ala Leu Asn Ser                        65     - #             70     - #             75                  - - GGA GCG CAG TCA CCC CAA GAG CAG CGC GAT GC - #A GGT ATC GTT GCC GCA          710                                                                       Gly Ala Gln Ser Pro Gln Glu Gln Arg Asp Al - #a Gly Ile Val Ala Ala                    80         - #         85         - #         90                      - - TCT GCA GGT AAC CAT GCC CAG GGC GTG GCC TA - #T GTG TGC AAG TCC TTG          758                                                                       Ser Ala Gly Asn His Ala Gln Gly Val Ala Ty - #r Val Cys Lys Ser Leu                95             - #    100             - #    105                          - - GGC GTT CAG GGA CGC ATC TAT GTT CCT GTG CA - #G ACT CCA AAG CAA AAG          806                                                                       Gly Val Gln Gly Arg Ile Tyr Val Pro Val Gl - #n Thr Pro Lys Gln Lys           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CGT GAC CGC ATC ATG GTT CAC GGC GGA GAG TT - #T GTC TCC TTG GTG        GTC      854                                                                    Arg Asp Arg Ile Met Val His Gly Gly Glu Ph - #e Val Ser Leu Val Val                          130  - #               135  - #               140              - - ACT GGC AAT AAC TTC GAC GAA GCA TCG GCT GC - #A GCG CAT GAA GAT GCA          902                                                                       Thr Gly Asn Asn Phe Asp Glu Ala Ser Ala Al - #a Ala His Glu Asp Ala                       145      - #           150      - #           155                  - - GAG CGC ACC GGC GCA ACG CTG ATC GAG CCT TT - #C GAT GCT CGC AAC ACC          950                                                                       Glu Arg Thr Gly Ala Thr Leu Ile Glu Pro Ph - #e Asp Ala Arg Asn Thr                   160          - #       165          - #       170                      - - GTC ATC GGT CAG GGC ACC GTG GCT GCT GAG AT - #C TTG TCG CAG CTG ACT          998                                                                       Val Ile Gly Gln Gly Thr Val Ala Ala Glu Il - #e Leu Ser Gln Leu Thr               175              - #   180              - #   185                          - - TCC ATG GGC AAG AGT GCA GAT CAC GTG GTG GT - #T CCA GTC GGC GGT GGC         1046                                                                       Ser Met Gly Lys Ser Ala Asp His Val Val Va - #l Pro Val Gly Gly Gly           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - GGA CTT CTT GCA GGT GTG GTC AGC TAC ATG GC - #T GAT ATG GCA CCT        CGC     1094                                                                    Gly Leu Leu Ala Gly Val Val Ser Tyr Met Al - #a Asp Met Ala Pro Arg                          210  - #               215  - #               220              - - ACT GCG ATC GTT GGT ATC GAA CCA GCG GGA GC - #A GCA TCC ATG CAG GCT         1142                                                                       Thr Ala Ile Val Gly Ile Glu Pro Ala Gly Al - #a Ala Ser Met Gln Ala                       225      - #           230      - #           235                  - - GCA TTG CAC AAT GGT GGA CCA ATC ACT TTG GA - #G ACT GTT GAT CCC TTT         1190                                                                       Ala Leu His Asn Gly Gly Pro Ile Thr Leu Gl - #u Thr Val Asp Pro Phe                   240          - #       245          - #       250                      - - GTG GAC GGC GCA GAG GTC AAA CGT GTC GGA GA - #T CTC AAC TAC ACC ATC         1238                                                                       Val Asp Gly Ala Glu Val Lys Arg Val Gly As - #p Leu Asn Tyr Thr Ile               255              - #   260              - #   265                          - - GTG GAG AAG AAC CAG GGT CGC GTG CGC ATG AT - #G AGC GCG ACC GAG GGC         1286                                                                       Val Glu Lys Asn Gln Gly Arg Val Arg Met Me - #t Ser Ala Thr Glu Gly           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GCT GTG TGT ACT GAG ATG CTC GAT CTT TAC CA - #A AAC GAA GGC ATC        ATC     1334                                                                    Ala Val Cys Thr Glu Met Leu Asp Leu Tyr Gl - #n Asn Glu Gly Ile Ile                          290  - #               295  - #               300              - - GCG GAG CCT GCT GGC GCG CTG TCT ATC GCT GG - #G TTG AAG GAA ATG TCC         1382                                                                       Ala Glu Pro Ala Gly Ala Leu Ser Ile Ala Gl - #y Leu Lys Glu Met Ser                       305      - #           310      - #           315                  - - TTT GCA CCT GGT TCT GTC GTG GTG TGC ATC AT - #C TCT GGT GGC AAC AAC         1430                                                                       Phe Ala Pro Gly Ser Val Val Val Cys Ile Il - #e Ser Gly Gly Asn Asn                   320          - #       325          - #       330                      - - GAT GTG CTG CGT TAT GCG GAA ATC GCT GAG CG - #C TCC TTG GTG CAC CGC         1478                                                                       Asp Val Leu Arg Tyr Ala Glu Ile Ala Glu Ar - #g Ser Leu Val His Arg               335              - #   340              - #   345                          - - GGT TCG AAG CAC TAC TTC TTG GTG AAC TTC CC - #G CAA AAG CCT GGT CAG         1526                                                                       Gly Ser Lys His Tyr Phe Leu Val Asn Phe Pr - #o Gln Lys Pro Gly Gln           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - TTG CGT CAC TTC CTG GAA GAT ATC CTG GGA CC - #G GAT GAT GAC ATC        ACG     1574                                                                    Leu Arg His Phe Leu Glu Asp Ile Leu Gly Pr - #o Asp Asp Asp Ile Thr                          370  - #               375  - #               380              - - CTG TTT GAG TAC CTC AAG CGC AAC AAC CGT GA - #G ACC GGT ACT GCG TTG         1622                                                                       Leu Phe Glu Tyr Leu Lys Arg Asn Asn Arg Gl - #u Thr Gly Thr Ala Leu                       385      - #           390      - #           395                  - - GTG GGT ATT CAC TTG AGT GAA GCA TCA GGA TT - #G GAT TCT TTG CTG GAA         1670                                                                       Val Gly Ile His Leu Ser Glu Ala Ser Gly Le - #u Asp Ser Leu Leu Glu                   400          - #       405          - #       410                      - - CGT ATG GAG GAA TCG GCA ATT GAT TCC CGT CG - #C CTC GAG CCG GGC ACT         1718                                                                       Arg Met Glu Glu Ser Ala Ile Asp Ser Arg Ar - #g Leu Glu Pro Gly Thr               415              - #   420              - #   425                          - - CCT GAG TAC GAA TAC TTG ACC TAAACATAGC TGAAGGCCA - #C CTCAATCGAG            1769                                                                       Pro Glu Tyr Glu Tyr Leu Thr                                                   430                 4 - #35                                                    - - GTGGCCTTTT TCTAGTTTCG GGTCAGGATC GCAAAGCCCC ACGGCTGAAG GG -             #TTGTGGAG   1829                                                                 - - GTGTCGGTGA CGGTGGGGGA AGTGAAGCTG TAAATCAGCT CGCCGCCAAG CG -            #GGACGGTG   1889                                                                 - - ATGGTGTCGT CGGAGAAATT CGCCAGAATT CGGCCG      - #                       - #     1925                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:12:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                              - - Met Ser Glu Thr Tyr Val Ser Glu Lys Ser Pr - #o Gly Val Met Ala        Ser                                                                               1               5 - #                 10 - #                 15             - - Gly Ala Glu Leu Ile Arg Ala Ala Asp Ile Gl - #n Thr Ala Gln Ala Arg                   20     - #             25     - #             30                  - - Ile Ser Ser Val Ile Ala Pro Thr Pro Leu Gl - #n Tyr Cys Pro Arg Leu               35         - #         40         - #         45                      - - Ser Glu Glu Thr Gly Ala Glu Ile Tyr Leu Ly - #s Arg Glu Asp Leu Gln           50             - #     55             - #     60                          - - Asp Val Arg Ser Tyr Lys Ile Arg Gly Ala Le - #u Asn Ser Gly Ala Gln       65                 - # 70                 - # 75                 - # 80       - - Ser Pro Gln Glu Gln Arg Asp Ala Gly Ile Va - #l Ala Ala Ser Ala Gly                       85 - #                 90 - #                 95              - - Asn His Ala Gln Gly Val Ala Tyr Val Cys Ly - #s Ser Leu Gly Val Gln                  100      - #           105      - #           110                  - - Gly Arg Ile Tyr Val Pro Val Gln Thr Pro Ly - #s Gln Lys Arg Asp Arg              115          - #       120          - #       125                      - - Ile Met Val His Gly Gly Glu Phe Val Ser Le - #u Val Val Thr Gly Asn          130              - #   135              - #   140                          - - Asn Phe Asp Glu Ala Ser Ala Ala Ala His Gl - #u Asp Ala Glu Arg Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ala Thr Leu Ile Glu Pro Phe Asp Ala Ar - #g Asn Thr Val Ile        Gly                                                                                             165  - #               170  - #               175             - - Gln Gly Thr Val Ala Ala Glu Ile Leu Ser Gl - #n Leu Thr Ser Met Gly                  180      - #           185      - #           190                  - - Lys Ser Ala Asp His Val Val Val Pro Val Gl - #y Gly Gly Gly Leu Leu              195          - #       200          - #       205                      - - Ala Gly Val Val Ser Tyr Met Ala Asp Met Al - #a Pro Arg Thr Ala Ile          210              - #   215              - #   220                          - - Val Gly Ile Glu Pro Ala Gly Ala Ala Ser Me - #t Gln Ala Ala Leu His      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Gly Gly Pro Ile Thr Leu Glu Thr Val As - #p Pro Phe Val Asp        Gly                                                                                             245  - #               250  - #               255             - - Ala Glu Val Lys Arg Val Gly Asp Leu Asn Ty - #r Thr Ile Val Glu Lys                  260      - #           265      - #           270                  - - Asn Gln Gly Arg Val Arg Met Met Ser Ala Th - #r Glu Gly Ala Val Cys              275          - #       280          - #       285                      - - Thr Glu Met Leu Asp Leu Tyr Gln Asn Glu Gl - #y Ile Ile Ala Glu Pro          290              - #   295              - #   300                          - - Ala Gly Ala Leu Ser Ile Ala Gly Leu Lys Gl - #u Met Ser Phe Ala Pro      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gly Ser Val Val Val Cys Ile Ile Ser Gly Gl - #y Asn Asn Asp Val        Leu                                                                                             325  - #               330  - #               335             - - Arg Tyr Ala Glu Ile Ala Glu Arg Ser Leu Va - #l His Arg Gly Ser Lys                  340      - #           345      - #           350                  - - His Tyr Phe Leu Val Asn Phe Pro Gln Lys Pr - #o Gly Gln Leu Arg His              355          - #       360          - #       365                      - - Phe Leu Glu Asp Ile Leu Gly Pro Asp Asp As - #p Ile Thr Leu Phe Glu          370              - #   375              - #   380                          - - Tyr Leu Lys Arg Asn Asn Arg Glu Thr Gly Th - #r Ala Leu Val Gly Ile      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - His Leu Ser Glu Ala Ser Gly Leu Asp Ser Le - #u Leu Glu Arg Met        Glu                                                                                             405  - #               410  - #               415             - - Glu Ser Ala Ile Asp Ser Arg Arg Leu Glu Pr - #o Gly Thr Pro Glu Tyr                  420      - #           425      - #           430                  - - Glu Tyr Leu Thr                                                                  435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:13:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 25 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                              - - CGCAGCTGAC TTCCATGGGG CAAGA          - #                  - #                   25                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:14:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                              - - CGTACGTTCG AACCGTGACC            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:15:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                              - - GGTCAGGGCA CCGTGGCTGC TG           - #                  - #                     22                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:16:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                              - - GGAGACTGTT GATCCCTTTG            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:17:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                              - - CCTTTGCACC TGGTTCTGTC            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:18:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                              - - CCTCAAGCGC AACAACCGTG AG           - #                  - #                     22                                                                    __________________________________________________________________________

We claim:
 1. A method for the microbial production of L-isoleucine,comprising(A) recombinantly preparing a microorganism having a mutatedthreonine dehydratase gene by replacing at least one base in the regionof the gene that encodes an allosteric domain of the threoninedehydratase, wherein said replacing at least one base results inderegulation of threonine dehydratase from L-isoleucine feedbackinhibition, thereby producing a recombinant microorganism, and (B)culturing the recombinant microorganism to produce L-isoleucine.
 2. Themethod of claim 1, wherein the microorganism is Corynebacteriumglutamicum.
 3. The method of claim 1, wherein the recombinantmicroorganism is obtained by growing the microorganism on a solid mediumwhich contains L-isoleucine and L-valine, and then selecting colonieswith an altered morphology.
 4. The method of claim 1, wherein saidmutated threonine dehydratase gene is obtained from Escherichia coli. 5.The method of claim 1, wherein said mutated threonine dehydratase geneis obtained from Corynebacterium glutamicum.
 6. The method of claim 1,wherein said replacing is selected from the group consisting of (i)replacing alanine in position 257 of SEQ ID NO: 8 with glycine and (ii)replacing methionine in position 199 of SEQ ID NO: 10 with valine. 7.The method of claim 1, wherein said replacing includes replacing, in SEQID NO:10, (a) histidine in position 278 with arginine and (b) leucine inposition 351 with serine.
 8. The method of claim 3, wherein acetohydroxyacid synthase activity of the microorganism has been inhibited by theL-valine contained in the solid medium.
 9. The method of claim 3,wherein the medium comprises a substance for inducing the threoninedehydratase gene.
 10. The method of claim 9, wherein said substance isisopropyl-β-D-thiogalactopyranoside.
 11. An isolated DNA that encodes athreonine dehydratase that is insensitive to L-isoleucine feedback, saidDNA prepared by replacing at least one base in the region of the DNAthat encodes an allosteric domain of the threonine-dehydratase.
 12. TheDNA of claim 11, comprising a nucleotide sequence selected from thegroup consisting of SEQ ID NO:1, SEQ ID NO:3, and SEQ ID NO:5.
 13. TheDNA of claim 11, comprising a nucleotide sequence selected from thegroup consisting of SEQ ID NO:7, SEQ ID NO:9, and SEQ ID NO:11.
 14. Acell comprising the DNA of claim
 11. 15. A vector comprising DNA thatencodes a threonine dehydratase that is insensitive to L-isoleucinefeedback, said DNA prepared by replacing at least one base in the regionof the DNA that encodes an allosteric domain of thethreonine-dehydratase.
 16. A vector according to claim 15, furthercomprising a promoter, inducible by isopropyl-β-D-thiogalactopyranoside,that is operably associated with said DNA.
 17. A cell comprising avector according to claim
 15. 18. A cell comprising a vector accordingto claim
 16. 19. The cell as claimed in claim 18, wherein said cellexhibits acetohydroxy acid synthase activity which can be inhibited byL-valine.
 20. The cell as claimed in claim 18, which is an Escherichiacoli cell.
 21. The cell as claimed in claim 18, which is aCorynebacterium glutamicum cell.
 22. The cell as claimed in claim 18,wherein said cell does not synthesize threonine dehydratase that issensitive to L-isoleucine feedback.